A Hollow Sphere Soft Lithography Approach for Long-Term Hanging Drop Methods
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Lee-2010-A Hollow Sphere Soft.pdf
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Author(s)
Lee, Won Gu
Ortmann, Daniel
Hancock, Matthew J.
Bae, Hojae
Khademhosseini, Ali
Date Issued
April 2010
Journal
Tissue Engineering. Part C, Methods
Publisher
Mary Ann Liebert
Citation
Lee, Won Gu et al. “A Hollow Sphere Soft Lithography Approach for Long-Term Hanging Drop Methods.” Tissue Engineering Part C: Methods 16.2 (2010): 249-259. Copyright © 2010, Mary Ann Liebert, Inc.
Version
Final published version
Abstract
In conventional hanging drop (HD) methods, embryonic stem cell aggregates or embryoid bodies (EBs) are often maintained in small inverted droplets. Gravity limits the volumes of these droplets to less than 50 μL [mu L], and hence such cell cultures can only be sustained for a few days without frequent media changes. Here we present a new approach to performing long-term HD methods (10–15 days) that can provide larger media reservoirs in a HD format to maintain more consistent culture media conditions. To implement this approach, we fabricated hollow sphere (HS) structures by injecting liquid drops into noncured poly(dimethylsiloxane) mixtures. These structures served as cell culture chambers with large media volumes (500 μL [mu L] in each sphere) where EBs could grow without media depletion. The results showed that the sizes of the EBs cultured in the HS structures in a long-term HD format were approximately twice those of conventional HD methods after 10 days in culture. Further, HS cultures showed multilineage differentiation, similar to EBs cultured in the HD method. Due to its ease of fabrication and enhanced features, this approach may be of potential benefit as a stem cell culture method for regenerative medicine.
MIT Department
Harvard University--MIT Division of Health Sciences and Technology
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1089/ten.tec.2009.0248
